草莓psy、pds和zds基因克隆及植物表达载体的构建  

Cloning of psy,pds and zds of Strawberry and Construction of Their Plant Expression Vectors

作　　者：朱海生[1,2,3] 陈敏氡[4] 林珲[1,2,3] 花秀凤[5] 温庆放[1,2,3] 

机构地区：[1]福建省农业科学院作物研究所,福建福州350013 [2]福建省农业科学院蔬菜研究中心,福建福州350013 [3]福建省蔬菜工程技术研究中心,福建福州350013 [4]福建农林大学园艺学院,福建福州350002 [5]福州市蔬菜科学研究所,福建福州350012

出　　处：《热带作物学报》2013年第1期54-60,共7页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金项目(No.31101534);福建省自然科学基金项目(No.2010J0111);福建省农业科学院科技创新团队重点科研项目(No.CXTD2011-20)

摘　　要：设计特异引物,分别克隆草莓类胡萝卜素合成关键基因psy、pds和zds开放阅读框,将psy和pds基因开放阅读框分别插入植物表达载体pBI121的CaMV 35S启动子和Nos终止子之间,构建了植物表达载体pBI121psy和pBI121pds。将psy和zds基因开放阅读框分别插入植物表达载体pCAMBIA1301的CaMV 35S启动子和Nos终止子之间,构建植物表达载体pCAMBIA1301psy和pCAMBIA1301zds。将带有完整启动子和终止子的pds基因引入pCAMBIA1301psy中,最终获得psy和pds的双价植物表达载体pCAMBIA1301psy-pds。经PCR、限制性内切酶酶切和测序鉴定后,成功将pBI121psy、pBI121pds、pCAMBIA1301psy、pCAMBIA1301zds和pCAMBIA1301psy-pds 5个重组表达质粒导入农杆菌EHA105中。该研究结果为进一步研究草莓psy、pds和zds基因的功能奠定基础。The open reading frame （ORF）of strawberry carotenoid biosynthesis genes psy, pals and zds were cloned, and the ORFs of the genes were inserted into the expression vectors pBI121 and pCAMBIAI30! between the CaMV 35S promoter and Nos terminator, and four plant vectors called pBI121psy, pBI121pds, pCAMBIA1301zds and pCAMBIA1301psy were obtained. The gene pds was also inserted into the expression vector pCAMBIA1301psy to generate double genes plant expression vector pCAMBIA1301psy-pds successfully. In addition, The identification results of PCI~ the restriction enzymes and DNA sequencing showed that the 5 recombinant expression plasmids pBI121psy, pBI121pds, pCAMBIA1301psy, pCAMBIA1301zds and pCAMBIA1301psy-pds were successfully introduced into Agrobacterium EHA105. The above research results layed a foundation for further studying the functions of psy, pds and zds genes.

关 键 词：草莓 psy PDS zds 表达载体 

分 类 号：Q943.2[生物学—植物学] Q949.751.8